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KR-20260066153-A - Modified nucleoside compound and oligonucleotide prepared therefrom

KR20260066153AKR 20260066153 AKR20260066153 AKR 20260066153AKR-20260066153-A

Abstract

As a nucleoside phosphoramidite compound, the compound is a compound with a 2'-phosphoramidite group and a 4'-modified TNA structure. By being incorporated into the head, end, and/or middle of an oligonucleotide, the modified oligonucleotide can hybridize with a portion of the target mRNA, thereby causing the normal function of the target mRNA to be lost or degraded.

Inventors

  • 장, 차오
  • 광, 솽
  • 위, 숴원
  • 황, 하오시
  • 수, 종하이

Assignees

  • 청두 브릴리언트 파마슈티컬 컴퍼니 리미티드
  • 아커나 리미티드

Dates

Publication Date
20260512
Application Date
20240925
Priority Date
20230925

Claims (20)

  1. As a nucleoside compound represented by formula (1), or a stereoisomer thereof, Here, R1 , R2 , R3 , and R4 are each independently selected from hydrogen, halogen, cycloalkyl group, C1-C6 alkyl group, substituted C1-C6 alkyl group, C1-C6 alkoxy group, substituted C1-C6 alkoxy group, C2-C6 alkenyl group, substituted C2-C6 alkenyl group, C2-C6 alkynyl group, or substituted C2-C6 alkynyl group; R 5 is selected from hydrogen, halogen, cyclopropane group, cyclobutane group, C1-C30 alkyl group, substituted C1-C30 alkyl group, C1-C30 alkoxy group, substituted C1-C30 alkoxy group, C2-C30 alkenyl group, substituted C2-C30 alkenyl group, C2-C30 alkynyl group or substituted C2-C30 alkynyl group; Nu indicates a base or its absence; Z is -O-, -S-, -NR 10 or -CR 10 R 11 , where R 10 and R 11 are each independently selected from hydrogen, halogen, acyl group, substituted or unsubstituted alkyl group, substituted or unsubstituted aryl group, substituted or unsubstituted heteroaryl group, substituted or unsubstituted heterocycle or substituted or unsubstituted cycloalkyl group; X 1 and X 2 are each independently selected from -(CH 2 ) n O- or -(CH 2 ) n S-, where n is any integer from 0 to 10; X 3 is selected from -(CH 2 ) n O-, -(CH 2 ) n -, -(CH 2 ) n S-, -(CH 2 ) n NH(CO)-, -(CH 2 ) n (CO)NH- or -(CH 2 ) n O(CH 2 ) n -A-, where A is a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted heterocyclic group, or a substituted or unsubstituted cycloalkyl group, where n is any integer from 0 to 10, a nucleoside compound or a stereoisomer thereof.
  2. In paragraph 1, The above nucleoside compound is a compound selected from Formula 1-a or 1-b, and: Here, R1 , R2 , R3 , R4 , R5 , Nu, X1 , X2 , and X3 are nucleoside compounds as defined in claim 1.
  3. In paragraph 2, Z is -O-, -S-, or -N(COR')-, where R' is a C1-C5 alkyl group; preferably, R' is selected from a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, and a tert-butyl group; X1 and X2 are each independently selected from -( CH2 ) nO- or -( CH2 ) nS- , where n is any integer from 0 to 5; preferably, n is selected from 0, 1, 2, 3, 4 or 5; X 3 is selected from -(CH 2 ) n -, -(CH 2 ) n O -, -(CH 2 ) n S -, -(CH 2 ) n NH(CO) -, -(CH 2 ) n (CO)NH - or -(CH 2 ) n O(CH 2 ) n -A -, wherein A is a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted heterocyclic group, or a substituted or unsubstituted cycloalkyl group, wherein n is any integer from 0 to 5; preferably, n is a nucleoside compound represented by Formula 1-a or 1-b selected from 0, 1, 2, 3, 4, or 5.
  4. In any one of paragraphs 1 through 3, Z is -O-, -S- or -N(COCH 3 )- and; X 1 is -O-, -S- or -CH 2 O-; X 2 is -O- or -S- and; X3 is -O-, -CH2- , -CH2O-, -CH2S- , -CH2NH (CO)- , -(CO)NH- or -CH2OCH2 -heteroaryl group-; preferably, X3 is -O-, -CH2O- , -CH2S- , -CH2NH (CO)-, -(CO)NH- or -CH2OCH2 - heteroaryl group-; R1 , R2 , and R3 are hydrogen; R 4 is hydrogen or a methoxy group; A nucleoside compound in which R5 is hydrogen, a methyl group, a halogen, or a C10-C18 alkyl group; preferably, R5 is a methyl group or a C10-C18 alkyl group; more preferably, R5 is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; and even more preferably, R5 is a methyl group, a C10 straight-chain alkyl group, a C11 straight-chain alkyl group, a C12 straight-chain alkyl group, a C13 straight-chain alkyl group, a C14 straight-chain alkyl group, a C15 straight-chain alkyl group, a C16 straight-chain alkyl group, a C17 straight-chain alkyl group, or a C18 straight-chain alkyl group.
  5. In any one of paragraphs 1 through 4, Nu is a base, and the base is a nucleoside compound selected from the following: .
  6. In any one of paragraphs 2 through 5, The nucleoside compound represented by Formula 1-a or Formula 1-b above is a nucleoside compound represented by Formula 2-a or Formula 2-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  7. In any one of paragraphs 2 through 5, The nucleoside compound represented by the above formula 1-a is selected from nucleoside compounds represented by formula 3-a, formula 5-a, formula 8-a, or formula 9-a, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a as defined in Claim 1.
  8. In any one of paragraphs 2 through 5, The nucleoside compound represented by Formula 1-a or Formula 1-b above is a nucleoside compound represented by Formula 4-a or Formula 3-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  9. In any one of paragraphs 2 through 5, The nucleoside compound represented by Formula 1-a or Formula 1-b above is a nucleoside compound represented by Formula 6-a or Formula 4-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  10. In any one of paragraphs 2 through 5, The nucleoside compound represented by Formula 1-a or Formula 1-b above is a nucleoside compound represented by Formula 7-a or Formula 5-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  11. In any one of paragraphs 2 through 5, The nucleoside compound represented by Formula 1-a or Formula 1-b above is a nucleoside compound represented by Formula 10-a or Formula 6-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  12. In any one of paragraphs 2 through 5, The nucleoside compound represented by Formula 1-a or Formula 1-b above is a nucleoside compound represented by Formula 11-a or Formula 7-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; R' is a C1-C5 alkyl group; preferably, R' is a methyl group, an ethyl group, an n-propyl group, or an isopropyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  13. In any one of paragraphs 2 through 5, The nucleoside compound represented by the above formula 1-a or formula 1-b is a nucleoside compound represented by formula 12-a or 8-b, respectively, and A nucleoside compound represented by Formula 1-a or Formula 1-b, wherein R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group.
  14. In any one of paragraphs 2 through 5, The nucleoside compound represented by the above Formula 1-a or Formula 1-b is a nucleoside compound represented by Formula 13-a or Formula 9-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  15. In any one of paragraphs 2 through 5, The nucleoside compound represented by Formula 1-a or Formula 1-b above is a nucleoside compound represented by Formula 14-a or Formula 10-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  16. In any one of paragraphs 2 through 5, The nucleoside compound represented by Formula 1-a or Formula 1-b above is a nucleoside compound represented by Formula 15-a or Formula 11-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  17. In any one of paragraphs 2 through 5, The nucleoside compound represented by the above formula 1-a or formula 1-b is a nucleoside compound represented by formula 16-a or 12-b, respectively, and R is a C1-C30 alkyl group, a substituted C1-C30 alkyl group, a C1-C30 alkoxy group, a substituted C1-C30 alkoxy group, a C2-C30 alkenyl group, a substituted C2-C30 alkenyl group, a C2-C30 alkynyl group, or a substituted C2-C30 alkynyl group; preferably, R is a methyl group, a C10 alkyl group, a C11 alkyl group, a C12 alkyl group, a C13 alkyl group, a C14 alkyl group, a C15 alkyl group, a C16 alkyl group, a C17 alkyl group, or a C18 alkyl group; Nu is a nucleoside compound represented by Formula 1-a or Formula 1-b as defined in claim 1.
  18. In paragraph 2 or 3, The nucleoside compound represented by the above formula 1-a is a nucleoside compound represented as follows: Here, Nu is a nucleoside compound represented by Formula 1-a as defined in Claim 1.
  19. In paragraph 2 or 3, The nucleoside compound represented by the above formula 1-a is a nucleoside compound represented by the following:
  20. In paragraph 2 or 3, The nucleoside compound represented by the above formula 1-b is a nucleoside compound represented as follows: Here, Nu is a nucleoside compound represented by Formula 1-b as defined in Claim 1.

Description

Modified nucleoside compound and oligonucleotide prepared therefrom The present invention relates to the field of biopharmaceuticals, and specifically to modified nucleoside compounds and oligonucleotides produced therefrom. Oligonucleotides are polymers of nucleotides. As nucleic acid inhibitor molecules, oligonucleotides can regulate intracellular mRNA levels, showing promising potential for the treatment of genetic diseases, metabolic diseases, cancer, and viral infections. Nucleic acid inhibitor molecules can regulate mRNA expression through various mechanisms, including RNA interference (RNAi). RNAi is a conserved pathway found in most eukaryotes, in which double-stranded RNA molecules (dsRNA) repress the expression of target genes having sequences complementary to the dsRNA. In a typical RNAi pathway, long dsRNA is cleaved by an endonuclease (Dicer) into short RNA double strands referred to as short interfering RNA ("siRNA"). siRNA binds to the endonuclease, trans-activating reaction RNA binding protein (TRBP), and Argonaute 2 ("Ago2") to form a complex referred to as the RNA-induced silencing complex ("RISC"). Ago2 is a nuclease that cleaves target mRNA by using the antisense strand (also referred to as the guide strand) of siRNA to induce sequence-specific cleavage of the target mRNA. Over the past few years, various double-stranded RNAi inhibitory molecular structures have been developed. For example, early work on RNAi inhibitory molecules focused on double-stranded nucleic acid molecules mimicking natural siRNA, each strand having 19 to 25 nucleotides and containing at least one 3' overhang having 1 to 5 nucleotides (see, e.g., U.S. Patent No. 8,372,968). Subsequently, long double-stranded RNAi inhibitory molecules were developed, which are cleaved in vivo by nucleases into active RNAi inhibitory molecules (see, e.g., U.S. Patent No. 8,883,996). In subsequent work, extended double-stranded nucleic acid inhibitor molecules were developed, wherein at least one strand extends beyond the double-stranded target region of said molecule, and said at least one strand comprises a thermodynamically stable tetracyclic structure (see, for example, U.S. Patent No. 8,513,207, U.S. Patent No. 8,927,705, WO2010/033225 and WO 2016/100401). Such structures include single-stranded extension structures (on one or both sides of said molecule) and double-stranded extension structures. In recent years, many modified nucleosides have been used in RNAi, including nucleoside modifications such as common 2'-fluorine modifications, 2'-methoxy group modifications, 2'-methoxyethyl group modifications, 2',3'-open ring nucleotides, lock nucleotides, ethylene glycol nucleotides, and 5'-(E)-vinyl phosphate. In some cases, chemical modifications are introduced into nucleoside inhibitor molecules to confer desirable properties under specific conditions, such as those occurring after in vivo administration. These modifications include designs to inhibit nucleases or other enzymes that interfere with the structure or activity of oligonucleotides, increase the cellular uptake of oligonucleotides, or improve the pharmacokinetic properties of oligonucleotides. For example, long-chain lipids are introduced into nucleosides (see, e.g., patent numbers WO2023283434, WO2020006050), and the modified nucleosides are introduced into the corresponding oligonucleotide sequences to improve the pharmacokinetic properties of oligonucleotide drugs and enhance their bioavailability in vivo. Further research on structural modifications of oligonucleotide drugs is required to obtain oligonucleotide drugs that are better in all respects. Figure 1 shows the remaining amount of the test sample at each time point in the 3'-exonuclease stability test of a single-stranded molecule in Experiment 4. Figure 2 shows the TTR protein inhibition rate of wild-type mice on day 21 of experiment 6. Figure 3 shows the liver exposure of mice on day 14 of experiment 7. Figure 4 shows the TTR protein inhibition rate of wild-type mice on day 21 of experiment 13. Figure 5 shows the results of the investigation into the TTR protein inhibitory activity of wild-type mice in Experiment 13. Unless otherwise specified, the apparatus used in the present invention is a conventional apparatus, and the reagents used are conventional reagents. The structure of a compound is determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). NMR shift (δ) is expressed in units of 10⁻⁶ (ppm). NMR measurements were performed using a Broker Avance III 400 NMR spectrometer, and the solvents used for the measurements were deuterium dimethyl sulfoxide (DMSO- d₆ ), deuterium chloroform ( CDCl₃ ), and deuterium methanol ( CD₃OD ), and the internal standard was tetramethylsilane (TMS); MS measurements were performed using (Agilent 6120B (ESI) and Agilent 6120B (APCI)); Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates are used for thin-layer chromatography, the dimensions of th